A Network Pharmacology-Based Investigation to the Pharmacodynamic Material Basis and Mechanisms of the Anti-Inflammatory and Anti-Viral Effect of Isatis indigotica

Abstract

Isatis indigotica (Ii) is a cruciferous herb that is widely distributed in China, and its roots and leaves have been used in two renowned antipyretic detoxicate crude drugs in Chinese Pharmacopoeia, Radix (R) and Folium (F) Isatidis. However, the pharmacodynamic material basis and underlying mechanisms of the herbal efficacy remained to be elucidated. Methods: Ultra-performance liquid chromatography-quadrupole-time-of-flight mass spectrometry (UPLC-Q-TOF-MS) was adopted for the chemical profiling of R and F Isatidis. The active ingredients were screened out through the prediction of gastrointestinal absorption and druglikeness analysis using SwissADME. A herb-ingredient-target network was constructed through target prediction of the herbal active ingredients and anti-inflammation or anti-viral properties, followed by protein-protein interaction analysis. Then, the potential relevant signaling pathways were predicted by pathway enrichment. Finally, for verification, RAW 264.7 cell line was adopted to examine the anti-inflammatory and anti-viral activities of 6 representative ingredients in Ii. Results: Seventy-three compounds have been identified from Ii through UPLC-Q-TOF-MS. A total of 17 potential active ingredients were screened through pharmacokinetics and druglikeness evaluation using SwissADME. It was shown that key targets might include TNF, AKT1, SRC, IL2, CASP9, and CASP3 in our herb-ingredient-target network, and isovitexin, a flavonoid, tended to participate in the inflammatory response, indoles were more likely to affect the cell proliferation processes, and lignans might have a broader affinity to key targets than the other active ingredients, such as regulating immune system (targeting IL-2) and PI3K-Akt signaling pathway. In vitro, indigo and secoisolariciresinol diglucoside markedly reduced TNF-α expression in Poly (I: C)-incubated cells. Isovitexin significantly inhibited TNF-α expression, and isatin treatment markedly reduced IL-1β expression in LPS-incubated cells. Conclusion:As the pharmacodynamics material basis of Ii, indoles, lignans, and flavonoids are believed to confer beneficial properties through various cellular aspects with multiple signaling pathways involved.

Computational Workflow to Study the Diversity of Secondary Metabolites in Fourteen Different Isatis Species

“…In addition, QC samples were employed every 10 injections to monitor the system stability. The mass spectrometer parameters were set according to our previous study with positive-ion as the acquisition mode [ 23 ].…”

“…In step 1, the AnalysisBaseFileConverter ( , accessed on 16 January 2022) software was used for raw mass spectrum data extraction, and MS-DIAL version 4.24 ( , accessed on 16 January 2022) was used for MS peak detection and alignment with the following parameters [ 10 ]: retention time range, 1–30 min (1–18 min for reference standard samples); retention time tolerance, 0.15 min; MS 1 mass range, 100–1200 (100–1000 min for reference standard samples); MS 1 tolerance, 0.01 Da; MS 2 mass range, 0–1200 (0–1000 min for reference standard samples); MS 2 tolerance, 0.02 Da; minimum peak height, 5000 amplitude (15,000 for reference standard samples). Other parameters, including mass slice width and MS/MS abundance cut off, were set according to our previous study [ 23 ]. An internal standard was used for peak heights normalization.…”

Anti-Inflammatory Effects of Phytochemical Components of Clinacanthus nutans

“…These effects are thought to occur via a decrease in the NF-κB pathway and an increase in the Nrf2-HO-1 pathway [ 65 ]. Isovitexin also inhibited TNF-α expression in LPS-incubated RAW 264.7 monocyte/macrophage-like cells [ 66 ].…”

Design, synthesis, and cytotoxic activities of isaindigotone derivatives as potential anti-gastric cancer agents

“…Chan’s research further implied that isaindigotone derivatives can downregulate c-myc transcription through disrupting the interaction of G-quardruplex and NM23-H2 20 . Recent study indicated that multiple signal pathways, including PI3K-AKT, were involved in the herbal efficacy of I. indigotica 22 . Same parent structure can also be found in PI3K delta inhibitor Idelalisib 23 .…”